Validation of the CTTv Fluorescent STR Multiplex System and the Amelogenin System
Sally Wallis, B.S.
Regional Forensic Science Center, Sedgwick County, Kansas, 1109 N. Minneapolis, Wichita,
Kansas 67214
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Validation studies were performed to determine the reproducibility, sensitivity, and specificity of the multiplex CTTv and Amelogenin systems, and to evaluate their applicability to forensic casework samples. Additionally, an optimal laboratory protocol was generated and compared to the protocol recommended by the Promega Corporation.
The multiplex contains the four STR loci CSF1PO, TPOX, TH01, vWA, and the sex-identification system Amelogenin. The GenePrintÔ CTTv (fluorescein-labeled) fluorescent STR kit and GenePrintÔ Sex Determination System-Amelogenin (fluorescein-labeled), both made by Promega Corporation, were utilized in these studies. The DNA was electrophoresed on the model SA-32 and bands were detected and analyzed using the Hitachi FMBIO® II.
The following areas were examined for validation: 1) inter- and intra-gel reproducibility of results; 2) specificity of results from body fluids other than blood; 3) human specificity; 4) determination of sensitivity of detection and sensitivity of extraction; 5) analysis of mixed stains; 6) inter- and intra-gel accuracy and precision of allelic ladders; and 7) the typing of biological samples from stimulated forensic conditions.
The optimal laboratory protocol generated was found to vary little from Promega’s recommended protocol. Furthermore, results of the studies indicate that STR typing using the multiplex system CTTv and the Amelogenin System is reliable, and these systems are applicable to forensic casework.
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